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Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro

The Sertoli cell tight junction (TJ) is the key component of the blood-testis barrier, where it sequesters developing germ cells undergoing spermatogenesis within the seminiferous tubules. Hormonally regulated claudin-11 is a critical transmembrane protein involved in barrier function and its murine...

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Autores principales: McCabe, Mark J, Foo, Caroline FH, Dinger, Marcel E, Smooker, Peter M, Stanton, Peter G
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Medknow Publications & Media Pvt Ltd 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4955190/
https://www.ncbi.nlm.nih.gov/pubmed/26585695
http://dx.doi.org/10.4103/1008-682X.163189
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author McCabe, Mark J
Foo, Caroline FH
Dinger, Marcel E
Smooker, Peter M
Stanton, Peter G
author_facet McCabe, Mark J
Foo, Caroline FH
Dinger, Marcel E
Smooker, Peter M
Stanton, Peter G
author_sort McCabe, Mark J
collection PubMed
description The Sertoli cell tight junction (TJ) is the key component of the blood-testis barrier, where it sequesters developing germ cells undergoing spermatogenesis within the seminiferous tubules. Hormonally regulated claudin-11 is a critical transmembrane protein involved in barrier function and its murine knockout results in infertility. We aimed to assess quantitatively the significance of the contribution of claudin-11 to TJ function, in vitro, using siRNA-mediated gene silencing. We also conducted an analysis of the contribution of occludin, another intrinsic transmembrane protein of the TJ. Silencing of claudin-11 and/or occludin was conducted using siRNA in an immature rat Sertoli cell culture model. Transepithelial electrical resistance was used to assess quantitatively TJ function throughout the culture. Two days after siRNA treatment, cells were fixed for immunocytochemical localization of junction proteins or lyzed for RT-PCR assessment of mRNA expression. Silencing of claudin-11, occludin, or both resulted in significant decreases in TJ function of 55% (P < 0.01), 51% (P < 0.01), and 62% (P < 0.01), respectively. Data were concomitant with significant decreases in mRNA expression and marked reductions in the localization of targeted proteins to the Sertoli cell TJ. We provide quantitative evidence that claudin-11 contributes significantly (P < 0.01) to Sertoli cell TJ function in vitro. Interestingly, occludin, which is hormonally regulated but not implicated in infertility until late adulthood, is also a significant (P < 0.01) contributor to barrier function. Our data are consistent with in vivo studies that clearly demonstrate a role for these proteins in maintaining normal TJ barrier structure and function.
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spelling pubmed-49551902016-07-26 Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro McCabe, Mark J Foo, Caroline FH Dinger, Marcel E Smooker, Peter M Stanton, Peter G Asian J Androl Original Article The Sertoli cell tight junction (TJ) is the key component of the blood-testis barrier, where it sequesters developing germ cells undergoing spermatogenesis within the seminiferous tubules. Hormonally regulated claudin-11 is a critical transmembrane protein involved in barrier function and its murine knockout results in infertility. We aimed to assess quantitatively the significance of the contribution of claudin-11 to TJ function, in vitro, using siRNA-mediated gene silencing. We also conducted an analysis of the contribution of occludin, another intrinsic transmembrane protein of the TJ. Silencing of claudin-11 and/or occludin was conducted using siRNA in an immature rat Sertoli cell culture model. Transepithelial electrical resistance was used to assess quantitatively TJ function throughout the culture. Two days after siRNA treatment, cells were fixed for immunocytochemical localization of junction proteins or lyzed for RT-PCR assessment of mRNA expression. Silencing of claudin-11, occludin, or both resulted in significant decreases in TJ function of 55% (P < 0.01), 51% (P < 0.01), and 62% (P < 0.01), respectively. Data were concomitant with significant decreases in mRNA expression and marked reductions in the localization of targeted proteins to the Sertoli cell TJ. We provide quantitative evidence that claudin-11 contributes significantly (P < 0.01) to Sertoli cell TJ function in vitro. Interestingly, occludin, which is hormonally regulated but not implicated in infertility until late adulthood, is also a significant (P < 0.01) contributor to barrier function. Our data are consistent with in vivo studies that clearly demonstrate a role for these proteins in maintaining normal TJ barrier structure and function. Medknow Publications & Media Pvt Ltd 2016 2015-11-10 /pmc/articles/PMC4955190/ /pubmed/26585695 http://dx.doi.org/10.4103/1008-682X.163189 Text en Copyright: © Asian Journal of Andrology http://creativecommons.org/licenses/by-nc-sa/3.0 This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.
spellingShingle Original Article
McCabe, Mark J
Foo, Caroline FH
Dinger, Marcel E
Smooker, Peter M
Stanton, Peter G
Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro
title Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro
title_full Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro
title_fullStr Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro
title_full_unstemmed Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro
title_short Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro
title_sort claudin-11 and occludin are major contributors to sertoli cell tight junction function, in vitro
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4955190/
https://www.ncbi.nlm.nih.gov/pubmed/26585695
http://dx.doi.org/10.4103/1008-682X.163189
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